BACKGROUND: At present, many problems deserve to be solved before clinical utilization of adipose tissue-derived mesenchymal stem cells (ADMSCs) such as complete differentiation from ADMSCs into cardiomyocytes and ADMSCs with or without specific function of cardiomyocytes. It is of significance to solve the problems including how to elevate the differentiation rate of ADMSCs into cardiomyocytes and how to elevate the homing and survival rate after transplantation for clinical utilization of stem cells.OBJECTIVE: To observe the differentiation of ADMSCs into cardiomyocytes after in vitro culture and induction.DESIGN: Randomized controlled observation.SETTING: Laboratory of Cardiology, General Hospital of Chinese PLA.MATERIALS: Adipose tissue was collected from abdominal operative patients at Department of General Surgery of General Hospital of Chinese PLA with the agreement of patients. Iscove's Modified Dulbecco's Medium (IMDM), type Ⅰ collagenase, 5-azacytidine (5-aza) and polyclonal antibody of specific anti-myocardial Troponin T (TnT) were purchased from Hyclon company, Gibco company, Sigma and Fujian Maixin Biotechnology Company, respectively.Monoclone antibodies of CD44, CD45, CD34, HLA2DR and factor-Ⅷ, Desmin, anti-α-striated muscle actin and anti-myosin heavy chain (MHC) were bought from Beijing Zhongshan Golden Bridge Biotechnology Limited Corporation. DAB stain, reverse transcription-polymerase chain reaction (RT-PCR) kit and atrial natriuretic peptide (ANP) radioimmunity kit were purchased from Beijing Zhongshan Golden Bridge Biotechnology Limited Corporation,Invitrogen and Radioimmunity Research Institute of Science and Technology Development Center of General Hospital of Chinese PLA, respectively.METHODS: The experiment was performed at the Laboratory of Cardiology, General Hospital of Chinese PLA from March 2005 to April 2006. ADMSCs were isolated and cultured by digestion and attachment culture method. The third generation of cells were determined by immunocytochemical method for surface molecule CD44, CD13, CD105, CD45,CD34, HLA-DR, factor Ⅷ and induced by addition of 5-aza into culture medium. On the 7th, 14th, 21st and 28th days, gene of GATA4 and Nkx2.5 were tested by reverse transcription-polymerase chain reaction (RT-PCR), and the concentration of atrial natriuretic polypeptide (ANP) were also examined by radioimmunoassay.MAIN OUTCOME MEASURES: Determination of cell surface marker, gene of GATA4 and Nkx2.5, and ANP concentration.and factor Ⅷ negative for the cultured cells. After induction for 7 days with 5-aza, no cells expressed Desmin,α-Sarcomeric Actin, Myocin heavy chain (MHC) and TnT, most cells were positive stained for CD44. After induction for 14 days, small amounts of cells displayed positive for Desmin,α-Sarcomeric Actin and MHC but still negative for TnT,while partial cells expressed positive CD44. After induction for 21 days, most cells were positive for Desmin,α-Sarcomeric after induction and no significant difference was found compared with that at the 7th day [(0.022±0.01) ng/L (P＞0.05].ANP concentration was respectively (7.92±0.21) and (8.12±0.50) ng/L at the 21st day and the 28th day (P＞0.05), but there was significant difference compared with that at the 7th day (P＜0.05).

Acupuncture Therapy ; Adolescent ; Humans ; Male ; Superior Mesenteric Artery Syndrome ; therapy

[ABSTRACT]OBJECTIVE To summarize the clinical related factors and prognostic influence factors of perioperative pulmonary embolism of head and neck malignant tumor.METHODSFrom 2010-2014, our hospital carried out a total of 2736 cases of head and neck malignant tumor surgical operations, of which, 10 cases were clinically diagnosed as postoperative pulmonary embolism, retrospectively analyzed the process of clinical treatment of the patients of pulmonary embolism with head and neck malignant tumors, and summarized their etiological factors, clinical manifestations, diagnosis and treatment. RESULTS The preoperative period incidence of pulmonary embolism in patients with head and neck malignant tumor was 0.37%(10/2736). Their clinical manifestations were mainly of asthma and breathing difficulty, and 3 cases of asymptomatic hypoxemia. 8 cases of patients showed pulmonary artery and branch filling defects after pulmonary artery angiography (CTA), 7 cases of patients got cured and were discharged from the hospital after comprehensive treatment such as anticoagulation etc; 3 cases of patients died after emergency treatment. 2 cases of patients suffered cavity bleeding, and there was no anticoagulant drug adjustment.CONCLUSIONThe mortality of head and neck cancer patients with perioperative pulmonary embolism is high, and therefore, preventive measures and timely treatments are important to reduce the incidence of pulmonary embolism.

Dust ; Humans ; Iron ; Mining ; Noise, Occupational ; Occupational Exposure ; statistics & numerical data ; Vibration

Plasma viral RNA load is widely accepted as the most relevant parameter to assess the status and progression of Simian immunodeficiency virus (SIV) infections. To accurately measure RNA levels of the virus, a one-step fluorescent quantitative assay was established based on the SYBR green Real-time reverse transcription-polymerase chain reaction (RT-PCR). The lower detection limit of the assay was 10 copies per reaction for the virus. This method was successfully applied to quantify SIVmac251 and SIVmac239 viruses produced in CEM×174 cells. Additionally, the performance of the SYBR green RT-PCR was assessed in a SIVmac251 infected rhesus macaque. The result demonstrated that the method could detect as little as 215 copies per milliliter of plasma and the dynamic pattern of viral load was highly consistent with previous results. With regard to convenience, sensitivity and accuracy our assay represents a realistic alternative to both branched-chain DNA (b-DNA) assays or real-time PCR assays based on TaqMan probes.

Objective To explore the relationship between brain-derived neurotrophic factor (BDNF)gene polymorphisms and the susceptibility to pediatric epilepsy.Methods BDNF polymorphisms in 128 patients with pediatric epilepsy and 132 healthy controls were analyzed with polymerase chain reaction restriction and fragment length polymorphism (PCR-RFLP).Results There were significant differences between pediatric epilepsy and controls on genotype frequency of BDNF-270C/T (X2 =7.08,P =0.03 ).The CC genotype was positively associated with pediatric epilepsy (OR =3.91,95％CI =1.26 ～ 12.14).No differences in genotype or allele frequencies of the other polymorphisms were found between patients and controls.The frequencies of haplotypes did not show significant differences between patients and controls.Conclusion These findings support the hypothesis that BDNF-270C/T polymorphism may contribute to the risk of developing pediatric epilepsy.

Objective To investigate the relationship between the C677T polymor-phism of the methylenetetrahydrofolate reductase (MTHFR) gene/G448A polymorphism of the

Objective To investigate the effects of morphine on PTEN expression and NF-κB activity in human gastric carcinoma cell line MGC-803.Methods The human gastric cancer cell line MGC-803 was purchased from Cell Biology Research Institute,Chinese Academy of Sciences,and cultured in DMEM liquid culture medium.The cells were randomly divided into 2 groups(n = 6 each): control group and morphine group.The cells was exposed to 0.1 μmol/L morphine in morphine group.The apoptosis was assessed by flow cytometry after being incubated with morphine for 24 h.PTEN expression and NF-κB activity were detected using RT-PCR and Western blot.Results The apoptotic rate was significantly increased,PTEN expression was up-regulated and NF-κB activity was significantly decreased in morphine group compared with control group(P ＜ 0.05).Conclusion Morphine can promote the apoptosis in human gastric cancer cells by up-regulating PTEN expression and decreasing NF-κB activity.

Objective To investigate the effect of fentanyl on the viability of human gastric cancer cell line MGC-803. Methods The human gastric cancer cell line MGC-803 was purchased from Cell Biology Research Institute, Chinese Academy of Sciences, and cultured in DMEM liquid culture medium. The cells were seeded in 6-well or 96-well plates and divided into 3 groups (n = 60 wells each): group Ⅰ normal control (group C); group Ⅱ and Ⅲ were exposed to fentanyl 0.01 and 1.00 μmol/L respectively (group F1, F2). The viability of the cells was detected by MTT assay after being incubated with fentanyl for 12, 24, 36, 48, 60 and 72 h. The cell cycle progression and apoptosis were assessed by flow cytometry and the ulrastructure of the cells was examined with transmission electron microscope after being incubated with fentanyl for 24 h. The proliferation of the cells was determined by colony formation assay at 7 day of incubation with fentanyl. Results The viability and proliferation of the cells and the proportion of the cells in S phase were significantly lower, while the proportion of the cella in G2/M phase and the apoptotic rate were higher in group F1 and F2 than in group C but no significant difference was found between group F1 and F2. The nuclear evelope was intact, the nucleolus and chromosomes were clearly visible in group C, while in group F1 and F2 fregmentation of nuclear envelope and nucleolus, chromatin condensation and apoptotic bodies were observed in group F2. Conclusion Fentanyl can inhibit the viability of human gastric cancer cells by its pro-apoptosis inducing effect.

Objective To explore the experience of self-perceived burden in patients with chronic obstructive pulmonary disease (COPD) and provide evidence for clinical nursing interventions.Methods Indepth interviews were conducted among 17 patients with COPD using phenomenological method of qualitative research.Data were analyzed with the Colaizzi's method.Results 3 themes was identified:self-accusation and guilt,loss of dignity and powerlessness,concern and insecurity.Conclusions Nurses should take the experience of SPB in patients with COPD seriously and take the influence of SPB into account during the process of nursing interventions and evaluation.